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ELEONORA TRUZZI
Assegnista di ricerca Dipartimento di Scienze della Vita sede ex-Scienze Farmaceutiche
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Pubblicazioni
2024
- 1H NMR targeted and untargeted approaches for the authentication of milk chocolates
[Articolo su rivista]
Truzzi, Eleonora; Rossi, Maria Cecilia; Bertelli, Davide
abstract
2024
- Phytochemical and functional characterization of cultivated varieties of Morus alba L. fruits grown in Italy
[Articolo su rivista]
Truzzi, E.; Marchetti, L.; Gibertini, G.; Benvenuti, S.; Cappellozza, S.; Giovannini, D.; Saviane, A.; Sirri, S.; Pinetti, D.; Assirelli, A.; Bertelli, D.
abstract
: Morus alba L. fruits are considered functional foods with an important nutritional value for their high content of polyphenols. Therefore, the type and level of phytochemicals of the soroses from 13 M. alba cultivars grown in Italy were characterized due to the lack of data available about their nutraceutical properties. Mature M. alba fruits exhibited variable polyphenol, flavonoid, anthocyanin, proanthocyanins, and 1-deoxynojirimycin contents which resulted in different antioxidant and α-glucosidase inhibitory activities. Regression models built on UHPLC-HRMS results revealed a strong correlation between the expression of quercetin derivatives, cyanidin 3-O-glucoside, caffeoyl methyl quinates, and 5,5'-dehydrodivanillic acid, and the biological activity of each fruit. On another note, principal component analysis revealed that the quantity of caffeoyl/dicaffeoyl methyl quinate, caffeoylquinic acids, and quercetin derivatives decreased during ripening. The results on the compositional and functional characterization of mature M. alba fruits might improve their consumption and economic value in Italy.
2023
- Combination of Nanodelivery Systems and Constituents Derived from Novel Foods: A Comprehensive Review
[Articolo su rivista]
Truzzi, E.; Bertelli, D.; Bilia, A. R.; Vanti, G.; Maretti, E.; Leo, E.
abstract
: Novel Food is a new category of food, regulated by the European Union Directive No. 2015/2283. This latter norm defines a food as "Novel" if it was not used "for human consumption to a significant degree within the Union before the date of entry into force of that regulation, namely 15 May 1997". Recently, Novel Foods have received increased interest from researchers worldwide. In this sense, the key areas of interest are the discovery of new benefits for human health and the exploitation of these novel sources of materials in new fields of application. An emerging area in the pharmaceutical and medicinal fields is nanotechnology, which deals with the development of new delivery systems at a nanometric scale. In this context, this review aims to summarize the recent advances on the design and characterization of nanodelivery systems based on materials belonging to the Novel Food list, as well as on nanoceutical products formulated for delivering compounds derived from Novel Foods. Additionally, the safety hazard of using nanoparticles in food products, i.e., food supplements, has been discussed in view of the current European regulation, which considers nanomaterials as Novel Foods.
2023
- Effects of Lavender Essential Oil Inhalation on the Welfare and Meat Quality of Fattening Heavy Pigs Intended for Parma Ham Production
[Articolo su rivista]
Nannoni, Eleonora; Martelli, Giovanna; Scozzoli, Maurizio; Belperio, Simona; Buonaiuto, Giovanni; Vannetti, Niccolò Ian; Truzzi, Eleonora; Rossi, Enrico; Benvenuti, Stefania; Sardi, Luca
abstract
2023
- Effects of Melaleuca alternifolia Chell (Tea Tree) and Eucalyptus globulus Labill. Essential Oils on Antibiotic-Resistant Bacterial Biofilms
[Articolo su rivista]
Iseppi, Ramona; Mariani, Martina; Benvenuti, Stefania; Truzzi, Eleonora; Messi, Patrizia
abstract
2023
- Multivariate Statistical Models for the Authentication of Traditional Balsamic Vinegar of Modena and Balsamic Vinegar of Modena on 1H-NMR Data: Comparison of Targeted and Untargeted Approaches
[Articolo su rivista]
Truzzi, Eleonora; Marchetti, Lucia; Piazza, Danny Vincenzo; Bertelli, Davide
abstract
This work aimed to compare targeted and untargeted approaches based on NMR data for the construction of classification models for Traditional Balsamic Vinegar of Modena (TBVM) and Balsamic Vinegar of Modena (BVM). Their complexity in terms of composition makes the authentication of these products difficult, which requires the employment of several time-consuming analytical methods. Here, 1H-NMR spectroscopy was selected as the analytical method for the analysis of TVBM and BVM due to its rapidity and efficacy in food authentication. 1H-NMR spectra of old (>12 years) and extra-old (>25 years) TVBM and BVM (>60 days) and aged (>3 years) BVM were acquired, and targeted and untargeted approaches were used for building unsupervised and supervised multivariate statistical modes. Targeted and untargeted approaches were based on quantitative results of peculiar compounds present in vinegar obtained through qNMR, and all spectral variables, respectively. Several classification models were employed, and linear discriminant analysis (LDA) demonstrated sensitivity and specificity percentages higher than 85% for both approaches. The most important discriminating variables were glucose, fructose, and 5-hydroxymethylfurfural. The untargeted approach proved to be the most promising strategy for the construction of LDA models of authentication for TVBM and BVM due to its easier applicability, rapidity, and slightly higher predictive performance. The proposed method for authenticating TBVM and BVM could be employed by Italian producers for safeguarding their valuable products.
2023
- NMR-based analytical methods for quantifying boswellic acids in extracts employed for producing food supplements: comparison of 13C-qNMR and 1H-NMR/PLS-R methods
[Articolo su rivista]
Truzzi, E.; Piazza, D. V.; Rossi, M. C.; Benvenuti, S.; Bertelli, D.
abstract
2023
- Novel application of 1H NMR spectroscopy coupled with chemometrics for the authentication of dark chocolate
[Articolo su rivista]
Truzzi, Eleonora; Marchetti, Lucia; Fratagnoli, Arianna; Rossi, Maria Cecilia; Bertelli, Davide
abstract
The applicability of 1H NMR spectroscopy coupled with chemometric in the quality control of dark chocolate was investigated for the first time to detect cocoa-butter equivalents (CBEs) above the allowed limit by European regulation. Blends of chocolate-fats with CBEs in the range 0-50 % were prepared and analyzed by 1H NMR spectroscopy. Datasets composed of peaks' areas or spectral variables (fingerprinting) in glycerol region were tested for the creation of multivariate statistical models. Partial least-squares discriminant analysis (PLS-DA) and regression (PLS-R) methods were used to correctly identify the type of CBE and quantify its concentration respectively. The performances of the models created on the two datasets were evaluated in terms of chemo-metric indicators and compared. The robustness of models was investigated through the analysis of test sets and random permutation tests. Fingerprinting models revealed fruitful results in classifying and quantifying CBEs in blends demonstrating the applicability of NMR in chocolate quality control.
2023
- Sistema per il rilascio controllato di 1-desossinojirimicina
[Brevetto]
Truzzi, Eleonora; Marchetti, Lucia; Bertelli, Davide; Cappellozza, Silvia; Saviane, Alessio
abstract
2022
- Characterization and Valorization of the Agricultural Waste Obtained from Lavandula Steam Distillation for Its Reuse in the Food and Pharmaceutical Fields
[Articolo su rivista]
Truzzi, Eleonora; Chaouch, Mohamed Aymen; Rossi, Gaia; Tagliazucchi, Lorenzo; Bertelli, Davide; Benvenuti, Stefania
abstract
2022
- Design and physicochemical characterization of novel hybrid SLN-liposome nanocarriers for the smart co-delivery of two antitubercular drugs
[Articolo su rivista]
Truzzi, Eleonora; Capocefalo, Angela; Meneghetti, Fiorella; Maretti, Eleonora; Mori, Matteo; Iannuccelli, Valentina; Domenici, Fabio; Castellano, Carlo; Leo, Eliana
abstract
2022
- Design, Characterization, and In Vitro Assays on Muscle Cells of Endocannabinoid-like Molecule Loaded Lipid Nanoparticles for a Therapeutic Anti-Inflammatory Approach to Sarcopenia
[Articolo su rivista]
Maretti, Eleonora; Molinari, Susanna; Battini, Renata; Rustichelli, Cecilia; Truzzi, Eleonora; Iannuccelli, Valentina; Leo, Eliana
abstract
Inflammatory processes play a key role in the pathogenesis of sarcopenia owing to their effects on the balance between muscle protein breakdown and synthesis. Palmitoylethanolamide (PEA), an endocannabinoid-like molecule, has been well documented for its anti-inflammatory properties, suggesting its possible beneficial use to counteract sarcopenia. The promising therapeutic effects of PEA are, however, impaired by its poor bioavailability. In order to overcome this limitation, the present study focused on the encapsulation of PEA in solid lipid nanoparticles (PEA-SLNs) in a perspective of a systemic administration. PEA-SLNs were characterized for their physico-chemical properties as well as cytotoxicity and cell internalization capacity on C2C12 myoblast cells. Their size was approximately 250 nm and the encapsulation efficiency reached 90%. Differential scanning calorimetry analyses demonstrated the amorphous state of PEA in the inner SLN matrix, which improved PEA dissolution, as observed in the in vitro assays. Despite the high internalization capacity observed with the flow cytometer (values between 85 and 94% after 14 h of incubation), the Nile Red labeled PEA-SLNs showed practically no toxicity towards myoblasts. Confocal analysis showed the presence of SLNs in the cytoplasm and not in the nucleus. These results suggest the potentiality provided by PEA-SLNs to obtain an innovative and side-effect-free tool in the medical treatment of sarcopenia
2022
- In vitro bioactivity evaluation of mulberry leaf extracts as nutraceutical for the management of diabete mellitus
[Articolo su rivista]
Marchetti, Lucia; Truzzi, Eleonora; Frosi, Ilaria; Papetti, Adele; Cappellozza, Silvia; Saviane, Alessio; Pellati, Federica; Bertelli, Davide
abstract
2022
- Plant Biostimulants Increase the Agronomic Performance of Lavandin (Lavandula x intermedia) in Northern Apennine Range
[Articolo su rivista]
Caccialupi, Giovanni; Caradonia, Federica; Ronga, Domenico; BEN HASSINE, Mortadha; Truzzi, Eleonora; Benvenuti, Stefania; Francia, Enrico
abstract
Lavandin (Lavandula x intermedia) belongs to the Lamiaceae family and is a shrub cultivated in the Mediterranean region for essential oils used to produce cosmetic, aromatherapy, and pharmaceutical ingredients. Nowadays, interest in plant biostimulants is rising due to their ability to increase biomass production in a sustainable way. The scope of the present study was to assess the effects of two plant biostimulants, one based on hydrolyzed proteins (FITOSIM®) and one based on seaweed extracts (FITOSTIM ALGA®), on the cultivar Grosso for two consecutive years in three different farms, located in the Italian Tuscan-Emilian Apennine Mountains. A difference in the efficiency of treatments among farms was shown, probably due to the plant age. In 2020, FITOSTIM ALGA® increased inflorescence fresh weights (+35%), while FITOSTIM® and FITOSTIM ALGA® enhanced stem and total fresh weights (+23% and +22%, respectively) compared to the untreated control. In 2021, both treatments enhanced the fresh and dry weights of inflorescence (+47% and +38%, respectively), while FITOSTIM ALGA® also improved the total plant dry weights (+34%). The plant biostimulants did not affect the chemical composition of essential oils. Our results indicate plant biostimulants as a supplement for sustainable management practices, enhancing Lavandin’s performance in mountainous agricultural areas.
2022
- Potential Applications of Essential Oils for Environmental Sanitization and Antimicrobial Treatment of Intensive Livestock Infections
[Articolo su rivista]
Mariotti, Melinda; Lombardini, Giulia; Rizzo, Silvia; Scarafile, Donatella; Modesto, Monica; Truzzi, Eleonora; Benvenuti, Stefania; Elmi, Alberto; Bertocchi, Martina; Fiorentini, Laura; Gambi, Lorenzo; Scozzoli, Maurizio; Mattarelli, Paola
abstract
2022
- Rapid Classification and Recognition Method of the Species and Chemotypes of Essential Oils by ATR-FTIR Spectroscopy Coupled with Chemometrics
[Articolo su rivista]
Truzzi, E.; Durante, C.; Bertelli, D.; Catellani, B.; Pellacani, S.; Benvenuti, S.
abstract
In the present work, the applicability of attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, coupled with chemometric tools in recognizing essential oils (EOs) for routine control, was evaluated. EOs belonging to Mentha, Cymbopogon, and Lavandula families and to S. rosmarinus and T. vulgaris species were analyzed, and the performance of several untargeted approaches, based on the synergistic combination of ATR-FTIR and Partial Least Squares Discriminant Analysis (PLS-DA), was tested to classify the species and chemotypes. Different spectra pre-processing methods were employed, and the robustness of the built models was tested by means of a Receiver Operating Characteristic (ROC) curve and random permutations test. The application of these approaches revealed fruitful results in terms of sensitivity and specificity, highlighting the potentiality of ATR-FTIR and chemometrics techniques to be used as a sensitive, cost-effective, and rapid tool to differentiate EO samples according to their species and chemotype.
2022
- Use of biostimulants to increase biomass production in Lavandin (Lavandula x intermedia) cultivated in Tuscan-Emilian Apennines
[Poster]
Caccialupi, Giovanni; Caradonia, Federica; BEN HASSINE, Mortadha; Truzzi, Eleonora; Benvenuti, Stefania; Ronga, Domenico; Francia, Enrico
abstract
Lavandin Lavandula x intermedia is one the most important perennial aromatic shrub widespread in the Italian marginal areas and is mainly cultivated for essentials oils production. Nowadays, in rural areas, biostimulants are gaining interest as useful product to enhance the biomass production, following the concept of sustainable agriculture. The present work aimed to evaluate the effects on biomass production of two biostimulants on lavandin crop cv “Grosso.” The biostimulants evaluated were: FITOSIM® composed from hydrolysed animal epithelium and FITOSIM ALGA® formulated from brown marine algae. The trial was conducted in three farms located in the Tuscan-Emilian Apennine area (Northern Italy). The products were applied as foliar spray fertilization during the blooming phase (weekly, in two times) of the years 2020 and 2021. The crops were transplanted in three different years: at Preci farm in 2016, at Campazzo farm in 2013 and at Pedroni farm in 2009. In the 2020, results showed that FITOSIM ALGA® increased the fresh weight of the inflorescence, moreover FITOSIM® and FITOSIM ALGA® treatments enhanced the stem and total fresh weights compared to the control. In 2021, treatments enhanced the inflorescence and total fresh weights, and the dry weight of the inflorescence, while FITOSTIM ALGA improved only the total plant dry weigh. Furthermore, the results showed that the year of transplant influenced the agronomic performances of the investigated biostimulants. In conclusion, this work suggests the ability of the biostimulants to enhance the biomass production on lavandin crop. However, further investigation will be required to evaluate the most efficient number of treatments, the effect of the biostimulants on yield parameters and on the quality of the essential oils.
2021
- A novel qnmr application for the quantification of vegetable oils used as adulterants in essential oils
[Articolo su rivista]
Truzzi, E.; Marchetti, L.; Benvenuti, S.; Righi, V.; Rossi, M. C.; Gallo, V.; Bertelli, D.
abstract
Essential oils (EOs) are more and more frequently adulterated due to their wide usage and large profit, for this reason accurate and precise authentication techniques are essential. This work aims at the application of qNMR as a versatile tool for the quantification of vegetable oils potentially usable as adulterants or diluents in EOs. This approach is based on the quantification of both1H and13C glycerol backbone signals, which are actually present in each vegetable oil containing triglycerides. For the validation, binary mixtures of rosemary EO and corn oil (0.8–50%) were prepared. To verify the general feasibility of this technique, other different mixtures including lavender, citronella, orange and peanut, almond, sunflower, and soy seed oils were analyzed. The results showed that the efficacy of this approach does not depend on the specific combination of EO and vegetable oil, ensuring its versatility. The method was able to determine the adulterant, with a mean accuracy of 91.81 and 89.77% for calculations made on1H and13C spectra, respectively. The high precision and accuracy here observed, make1H-qNMR competitive with other well-established techniques. Considering the current importance of quality control of EOs to avoid fraudulent practices, this work can be considered pioneering and promising.
2021
- Attenuated total reflectance–Fourier transform infrared (ATR–FTIR) spectroscopy coupled with chemometric analysis for detection and quantification of adulteration in lavender and citronella essential oils
[Articolo su rivista]
Truzzi, E.; Marchetti, L.; Bertelli, D.; Benvenuti, S.
abstract
Introduction: The growing consumer interest in “naturals” led to an increased application of essential oils (EOs). The market outbreak induced the intensification of EO adulterations, which could affect their quality. Objectives: Nowadays, little is known about the illegal practice of adulteration of EOs with vegetable oils. Therefore, the application of mid-infrared spectroscopy coupled with chemometrics was proposed for the detection of EO counterfeits. Materials and methods: Two EOs, three seed oils, and their mixtures were selected to build the adulteration model. EO–adulterant mixtures for model calibration and validation were analyzed by attenuated total reflectance–Fourier transform infrared (ATR–FTIR) spectroscopy. The spectral data were analyzed with principal component analysis (PCA) and partial least-squares (PLS) regression. Results: PCA allowed the discrimination of the EO and adulterant percentages by explaining 97.47% of the total spectral variance with two principal components. A PLS regression model was generated with three factors explaining 97.73% and 99.69% of the total variance in X and Y, respectively. The root mean square error of calibration and the root mean square error of cross-validation were 0.918 and 1.049, respectively. The root mean square error of prediction value obtained from the external validation set was 1.588 and the coefficients of determination R2CAL and R2CV were 0.997 and 0.996, respectively. Conclusions: The results highlighted the robustness of the developed method in quantifying counterfeits in the range from 0 to 50% of adulterants, disregarding the type of EO and adulterant employed. The present work offers a research advance and makes an important impact in phytochemistry, revealing an easily applicable method for EO quality assessment.
2021
- Development of solid lipid nanoparticles for the delivery of an anti- inflammatory drug to muscle skeletal cells
[Poster]
Maretti, Eleonora; Truzzi, Eleonora; Molinari, Susanna; Battini, Renata; Leo, Eliana Grazia
abstract
2021
- Drug delivery strategies of Geraniol via nose-to-brain for the treatment of neuronal disorders
[Abstract in Atti di Convegno]
Truzzi, Eleonora; Dalpiaz, Alessandro; Rustichelli, Cecilia; Ferraro, Luca; de Oliveira Junior Edilson, Ribeiro; Fogagnolo, Marco; Pavan, Barbara; Beggiato, Sarah; Lima, Eliana; Leo, Eliana
abstract
Parkinson’s disease (PD) is a neurodegenerative disorder which affects 1% of people over 65 years. Neuroinflammation and mitochondrial dysfunction play a central role in the disorder progression. Therefore, anti-oxidants, promoters of neurotrophic factor expression and mitochondrial rescuers could prevent the disease progression. Geraniol (GER) is a natural compound with demonstrated anti-oxidant and neuroprotective activities in PD models. However, its activity is impaired by a fast metabolism following oral administration, with a half-life of about 12 minutes. To overcome this limitation, nose-to-brain administration is a promising strategy because it allows the direct delivery of drugs beyond the blood-brain barrier avoiding the first-pass metabolism. However, GER is highly volatile and irritant. Thus, an appropriate delivery system for its nose-to-brain administration is required. Two different strategies were developed in order to achieve a successful nose to brain delivery. The first one aimed the encapsulation of pure GER and GER prodrug in solid lipid nanoparticles (SLNs), while the second one, displayed here, yearns to complex GER with cyclodextrins (CDs). The inclusion complexes were designed in order to reduce GER volatility and to obtain long-term stable formulations as powders.
2021
- In vivo β-carotene skin permeation modulated by Nanostructured Lipid Carriers
[Articolo su rivista]
Maretti, E.; Leo, E.; Rustichelli, C.; Truzzi, E.; Siligardi, C.; Iannuccelli, V.
abstract
Nanostructured Lipid Carriers (NLC) were investigated with the purpose of promoting skin permeation of the highly lipophilic β-carotene (BC) across the stratum corneum (SC) barrier so that it may perform its antioxidant properties in photo-aging and epithelial skin cancer prevention. Two differently sized NLC samples were developed using stearic acid and squalene as lipid matrix and evaluated in comparison with Microstructured Lipid Carriers (MLC). The carriers were characterized for morphology, size, Z-potential, BC loading and release as well as physical state by means of DSC and XRPD analyses. In vivo penetration of the carriers was assessed on humans by determining BC concentrations within the SC stratum disjunctum and stratum compactum layers removed by means of the tape stripping test in comparison with pure BC. Unlike MLC and pure BC that were mostly retained within the outermost layers of the SC, the NLC sample having the smallest size (about 200 nm) has proved to penetrate more deeply into the SC barrier. Accordingly, the goal of providing β-carotene actions against oxidative damages within the looser skin viable tissues could be envisaged.
2021
- Nasal biocompatible powder of geraniol oil complexed with cyclodextrins for neurodegenerative diseases: Physicochemical characterization and in vivo evidences of nose to brain delivery
[Articolo su rivista]
Truzzi, Eleonora; Rustichelli, Cecilia; de Oliveira Junior, Edilson Ribeiro; Ferraro, Luca; Maretti, Eleonora; Graziani, Daniel; Botti, Giada; Beggiato, Sarah; Iannuccelli, Valentina; Lima, Eliana Martins; Dalpiaz, Alessandro; Leo, Eliana
abstract
Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-inflammatory effects in neurodegenerative diseases and, in particular, Parkinson's disease (PD) models. Unfortunately, delivering GER to the CNS via nose-to-brain is not feasible due to its irritant effects on the mucosae. Therefore, in the present study β-cyclodextrin (βCD) and its hydrophilic derivative hydroxypropyl-beta-cyclodextrin (HPβCD) were selected as potential carriers for GER nose-to-brain delivery. Inclusion complexes were formulated and the biocompatibility with nasal mucosae and drug bioavailability into cerebrospinal fluid (CSF) were studied in rats. It has been demonstrated by DTA, FT-IR and NMR analyses that both the CDs were able to form 1:1 GER-CD complexes, arising long-term stable powders after the freeze-drying process. GER-HPβCD-5 and GER-βCD-2 complexes exhibited comparable results, except for morphology and solubility, as demonstrated by SEM analysis and phase solubility study, respectively. Even though both complexes were able to directly and safely deliver GER to CNS, GER-βCD-2 displayed higher ability in releasing GER in the CSF. In conclusion, βCD complexes can be considered a very promising tool in delivering GER into the CNS via nose-to-brain route, preventing GER release into the bloodstream and ensuring the integrity of the nasal mucosa.
2021
- Novel Strategy for the Recognition of Adulterant Vegetable Oils in Essential Oils Commonly Used in Food Industries by Applying 13C NMR Spectroscopy
[Articolo su rivista]
Truzzi, Eleonora; Marchetti, Lucia; Benvenuti, Stefania; Ferroni, Annalisa; Rossi, Maria Cecilia; Bertelli, Davide
abstract
Essential oils (EOs) are valuable products commonly employed in the food industry and intensively studied as biopreservatives for the extension of food shelf-life. Unfortunately, EOs might be counterfeit to increase industrial profits. Among the possible adulterants, vegetable oils (VOs) must be considered for their characteristics and low costs. We aimed to apply nuclear magnetic resonance (NMR) spectroscopy for the detection and identification of VOs in mixtures with EOs. This innovative strategy is based on comparing the peak area ratio matrices of characteristic VO 13C NMR fatty acid signals with those of adulterated EOs. The identification of the VOs was achieved by calculating the matrix similarity at different confidence levels. The strategy demonstrated the capacity to efficiently recognize the presence of adulteration and the type of VO adulterant in mixtures. Thus, the method was applied to 20 commercial EOs, and VOs were detected and then identified in four samples.
2020
- In Vivo Biodistribution of Respirable Solid Lipid Nanoparticles Surface-Decorated with a Mannose-Based Surfactant: A Promising Tool for Pulmonary Tuberculosis Treatment?
[Articolo su rivista]
Truzzi, Eleonora; Leite Nascimento, Thais; Iannuccelli, Valentina; Costantino, Luca; Martins Lima, Eliana; Leo, Eliana Grazia; Siligardi, Cristina; Gualtieri, Eva Magdalena; Maretti, Eleonora
abstract
The active targeting to alveolar macrophages (AM) is an attractive strategy to improve the therapeutic efficacy of ‘old’ drugs currently used in clinical practice for the treatment of pulmonary tuberculosis. Previous studies highlighted the ability of respirable solid lipid nanoparticle assemblies (SLNas), loaded with rifampicin (RIF) and functionalized with a novel synthesized mannose-based surfactant (MS), both alone and in a blend with sodium taurocholate, to efficiently target the AM via mannose receptor-mediated mechanism. Here, we present the in vivo biodistribution of these mannosylated SLNas, in comparison with the behavior of both non-functionalized SLNas and bare RIF. SLNas biodistribution was assessed, after intratracheal instillation in mice, by whole-body real-time fluorescence imaging in living animals and RIF quantification in excised organs and plasma. Additionally, SLNas cell uptake was determined by using fluorescence microscopy on AM from bronchoalveolar lavage fluid and alveolar epithelium from lung dissections. Finally, histopathological evaluation was performed on lungs 24 h after administration. SLNas functionalized with MS alone generated the highest retention in lungs associated with a poor spreading in extra-pulmonary regions. This effect could be probably due to a greater AM phagocytosis with respect to SLNas devoid of mannose on their surface. The results obtained pointed out the unique ability of the nanoparticle surface decoration to provide a potential more efficient treatment restricted to the lungs where the primary tuberculosis infection is located.
2020
- Nasal administration of nanoencapsulated geraniol/ursodeoxycholic acid conjugate: Towards a new approach for the management of Parkinson's disease
[Articolo su rivista]
de Oliveira Junior, Edilson Ribeiro; Truzzi, Eleonora; Ferraro, Luca; Fogagnolo, Marco; Pavan, Barbara; Beggiato, Sarah; Rustichelli, Cecilia; Maretti, Eleonora; Lima, Eliana Martins; Leo, Eliana; Dalpiaz, Alessandro
abstract
The combined use of different therapeutic agents in the treatment of neurodegenerative disorders is a promising strategy to halt the disease progression. In this context, we aimed to combine the anti-inflammatory properties of geraniol (GER) with the mitochondrial rescue effects of ursodeoxycholic acid (UDCA) in a newly-synthesized prodrug, GER-UDCA, a potential candidate against Parkinson's disease (PD). GER-UDCA was successfully synthetized and characterized in vitro for its ability to release the active compounds in physiological environments. Because of its very poor solubility, GER-UDCA was entrapped into both lipid (SLNs) and polymeric (NPs) nanoparticles in order to explore nose-to-brain pathway towards brain targeting. Both GER-UDCA nanocarriers displayed size below 200 nm, negative zeta potential and the ability to increase the aqueous dissolution rate of the prodrug. As SLNs exhibited the higher GER-UDCA dissolution rate, this formulation was selected for the in vivo GER-UDCA brain targeting experiments. The nasal administration of GER-UDCA-SLNs (1 mg/kg of GER-UDCA) allowed to detect the prodrug in rat cerebrospinal fluid (concentration range = 1.1 to 4.65 μg/mL, 30–150 min after the administration), but not in the bloodstream, thus suggesting the direct nose to brain delivery of the prodrug. Finally, histopathological evaluation demonstrated that, in contrast to the pure GER, nasal administration of GER-UDCA-SLNs did not damage the structural integrity of the nasal mucosa. In conclusion, the present data suggest that GER-UDCA-SLNs could provide an effective and non-invasive approach to boost the access of GER and UDCA to the brain with low dosages.
2020
- Physicochemical characterization of SLN/Liposomes hybrid nanoparticles for the co-delivery of two antitubercular drugs: focus on SANS analysis
[Poster]
Truzzi, Eleonora; Meneghetti, Fiorella; Mori, Matteo; Maretti, Eleonora; Domenici, Fabio; Castellano, Carlo; Capocefalo, Angela; Leo, Eliana Grazia
abstract
2019
- Drugs/lamellae interface influences the inner structure of double-loaded liposomes for inhaled anti-TB therapy: An in-depth small-angle neutron scattering investigation
[Articolo su rivista]
Truzzi, Eleonora; Meneghetti, Fiorella; Mori, Matteo; Costantino, Luca; Iannuccelli, Valentina; Maretti, Eleonora; Domenici, Fabio; Castellano, Carlo; Rogers, Sarah; Capocefalo, Angela; Leo, Eliana
abstract
With the aim of developing new drug carriers for inhalation therapy, we report here an in depth investigation of the structure of multilamellar liposomes loaded with two well-established anti-tubercular (anti-TB) drugs, isoniazid (INH) and rifampicin (RIF), by means of small-angle neutron-scattering (SANS) analysis. Unloaded, single drug-loaded and co-loaded liposomes were prepared using different amounts of drugs and characterized regarding size, encapsulation efficiency and drug release. Detailed information on relevant properties of the investigated host-guest structures, namely the steric bilayer thickness, particle dispersion, number of lamellae and drug localization was studied by SANS. Results showed that RIF-liposomes were less ordered than unloaded liposomes. INH induced a change in the inter-bilayer periodical spacing, while RIF-INH co-loading stabilized the multilamellar liposome architecture, as confirmed by the increment of the drug loading capacity. These findings could be useful for the understanding of in vitro and in vivo behavior of these systems and for the design of new drug carriers, intended for inhaled therapy.
2019
- Newly synthesized surfactants for surface mannosylation of respirable SLN assemblies to target macrophages in tuberculosis therapy
[Articolo su rivista]
Maretti, Eleonora; Costantino, Luca; Buttini, Francesca; Rustichelli, Cecilia; Leo, Eliana Grazia; Truzzi, Eleonora; Iannuccelli, Valentina
abstract
The present study reports about new Solid Lipid Nanoparticle assemblies (SLNas) loaded with rifampicin (RIF) surface-decorated with novel mannose derivatives, designed for anti-tuberculosis (TB) inhaled therapy by dry powder inhaler (DPI). Mannose is considered a relevant ligand to achieve active drug targeting being mannose receptors (MR) overexpressed on membranes of infected alveolar macrophages (AM), which are the preferred site of Mycobacterium tuberculosis. Surface decoration of SLNas was obtained by means of newly synthesized functionalizing compounds used as surfactants in the preparation of carriers. SLNas were fully characterized in vitro determining size, morphology, drug loading, drug release, surface mannosylation, cytotoxicity, macrophage internalization extent and ability to bind MR, and intracellular RIF concentration. Moreover, the influence of these new surface functionalizing agents on SLNas aerodynamic performance was assessed by measuring particle respirability features using Next Generation Impactor. SLNas exhibited suitable drug payload, in vitro release, and more efficient ability to enter macrophages (about 80%) compared to bare RIF (about 20%) and to non-functionalized SLNas (about 40%). The involvement of MR specific binding has been demonstrated by saturating MR of J774 cells causing a decrease of RIF intracellular concentration of about 40%. Furthermore, it is noteworthy that the surface-decoration of particles produced a poor cohesive powder with an adequate respirability (fine particle fraction ranging from about 30% to 50%). Therefore, the proposed SLNas may represent an encouraging opportunity in a perspective of an efficacious anti-TB inhaled therapy.
2019
- Novel engineered lipid-based nanoparticles for pulmonary tuberculosis inhalation therapy
[Poster]
Maretti, Eleonora; Truzzi, Eleonora; Costantino, Luca; Rustichelli, Cecilia; Martins Lima, Eliana; Leite Nascimento, Thais; Siligardi, Cristina; Gualtieri, Eva Magdalena; Miselli, Paola; Buttini, Francesca; Leo, Eliana Grazia; Iannuccelli, Valentina
abstract
Priorities to achieve the WHO goal of ending tuberculosis (TB) epidemic by 2030 include new drug treatments to simplify and shorter conventional drug regimens. TB is caused by Mycobacterium tuberculosis residing and surviving inside alveolar macrophages (AM). Considering that 75-80% of cases of infection remain localized in the lungs, the easiest and most successful therapy could involve the inhalation route offering benefits in terms of patient’s autonomy and compliance, by-passing hepatic metabolism, reducing dose amount, dose frequency, and treatment duration, thus minimising the risk of drug-resistant mutants, toxicity, and side effects. Inhalable powder formulations of repurposed drugs entail engineering techniques such as micro- or nanoparticulate carriers enabling drug emission by Dry Powder Inhaler devices, deposition onto alveolar epithelia, and transport into AM. Within this context, Solid Lipid Nanoparticle assemblies (SLNas) loaded with rifampicin, a clinically useful anti-TB drug, were produced by processing accepted excipients for DPI formulations through an optimized methodology that avoids organic solvents and is suitable for a large-scale production. The prototypes were functionalized by means of newly synthesized AM receptor-specific targeting agents as the ligands anchored on SLNas surface without chemical reactions. In vitro and in vivo preclinical studies highlighted functionalized SLNas with adequate respirability performance, safety, AM internalization ability, and mice lung deposition in an encouraging perspective of a potential efficacious pulmonary TB therapy. This research was supported by a grant on the project “FAR interdisciplinare 2017” from the University of Modena and Reggio Emilia, Modena, Italy (PI Prof. Luca Costantino)
2019
- Self-assembled organogelators as artificial stratum corneum models: key-role parameters for skin permeation prediction
[Articolo su rivista]
Maretti, Eleonora; Rustichelli, Cecilia; Miselli, Paola; Leo, Eliana Grazia; Truzzi, Eleonora; Iannuccelli, Valentina
abstract
Self-assembled organogelators were explored as artificial stratum corneum (SC) models for the in vitro skin permeation assessment. Four SC models consisting of binary (organogels) or ternary (microemulsion-based organogels) mixtures were developed using stearic acid, tristearin, or sorbitan tristearate, at two different concentrations, gelled in squalene. The permeation of lipophilic butyl-methoxydibenzoylmethane and hydrophilic methylene blue as the permeant compounds across the SC models was compared with ex vivo experiments using excised porcine ear skin. A multi-analytical approach was adopted to provide detailed understanding about organogelator organization within the SC models and find possible parameters playing key-roles in SC permeation prediction. The SC models were investigated for gelling properties and microstructure. Parameters such as gel occurrence, organogelator concentration, and rheological properties appeared as negligible conditions for skin permeation prediction. Conversely, arrangement packing, interactions, and crystallinity extent of the self-assembled organogelator were found to play a fundamental role in the simulation of SC barrier function according to the permeant feature.
2019
- The Impact of Lipid Corona on Rifampicin Intramacrophagic Transport Using Inhaled Solid Lipid Nanoparticles Surface-Decorated with a Mannosylated Surfactant
[Articolo su rivista]
Maretti, Eleonora; Rustichelli, Cecilia; Gualtieri, Eva Magdalena; Costantino, Luca; Siligardi, Cristina; Miselli, Paola; Buttini, Francesca; Montecchi, Monica; Leo, Eliana Grazia; Truzzi, Eleonora; Iannuccelli, Valentina
abstract
The mimicking of physiological conditions is crucial for the success of accurate in vitro
studies. For inhaled nanoparticles, which are designed for being deposited on alveolar epithelium
and taken up by macrophages, it is relevant to investigate the interactions with pulmonary
surfactant lining alveoli. As a matter of fact, the formation of a lipid corona layer around the
nanoparticles could modulate the cell internalization and the fate of the transported drugs. Based
on this concept, the present research focused on the interactions between pulmonary surfactant and
Solid Lipid Nanoparticle assemblies (SLNas), loaded with rifampicin, an anti-tuberculosis drug.
SLNas were functionalized with a synthesized mannosylated surfactant, both alone and in a blend
with sodium taurocholate, to achieve an active targeting to mannose receptors present on alveolar
macrophages (AM). Physico-chemical properties of the mannosylated SLNas satisfied the
requirements relative to suitable respirability, drug payload, and AM active targeting. Our studies
have shown that a lipid corona is formed around SLNas in the presence of Curosurf, a commercial
substitute of the natural pulmonary surfactant. The lipid corona promoted an additional resistance
to the drug diffusion for SLNas functionalized with the mannosylated surfactant and this improved
drug retention within SLNas before AM phagocytosis takes place. Moreover, lipid corona formation
did not modify the role of nanoparticle mannosylation towards the specific receptors on MH-S cell
membrane.
2018
- Organo-modified bentonite for gentamicin topical application: interlayer structure and in vivo skin permeation
[Articolo su rivista]
Iannuccelli, Valentina; Maretti, Eleonora; Bellini, Alessia; Malferrari, Daniele; Ori, Guido; Montorsi, Monia; Bondi, Moreno; Truzzi, Eleonora; Leo, Eliana Grazia
abstract
Recent biomedical applications of clay materials have included organically modified clays or clay minerals with the purpose of modifying and improving drug biological activity. The present research aims to explore the potential benefits provided by a raw bentonite (Bt) modified by gentamicin (GM) adsorbed within montmorillonite interlayers in the management of cutaneous infectious diseases. Information arisen from controlled X-ray powder diffraction, thermogravimetry coupled with evolved gas mass spectrometry, and molecular dynamics simulations pointed out GM monolayer arrangement within montmorillonite framework without producing substantial effects on the layer periodicity. Concerning skin biomedical application, unlike the pure antibiotic permeating along the trans-follicular pathway across stratum corneum, the organo-modified Bt/GM would favor the trans-epidermal route along inter-cluster corneocyte region, as in vivo skin penetration studies by means of tape stripping test indicated. Based on the results obtained, GM intercalation could represent a potential advantageous approach allowing a long-term Bt/GM reservoir for sustained antibacterial activity.
2018
- Polymeric and Solid Lipid Nanoparticles for nose-to-brain delivery of geraniolursodeoxycholic
acid conjugate: development and characterization studies
[Abstract in Atti di Convegno]
Edilson Oliveira Junior, ; Truzzi, Eleonora; Maretti, Eleonora; Leo, Eliana Grazia; Dalpiaz, Alessandro; Rustichelli, Cecilia; Marco, Fogagnolo; Eliana, Lima
abstract
Neurodegenerative disorder treatment is a challenge mainly due to the difficulty of drug transport across the
blood-brain barrier [1]. Intranasal administration of nanoparticles as carrier system may increase drug
concentration into the brain [2]. Geraniol (GER) has demonstrated antioxidant and neuroprotective activities in
Parkinson’s disease animal models [3]. However, due to its volatility, GER is hardly incorporated into freezedrying
nanoparticles. On the other hand, GER-ursodeoxycholic acid conjugate (GER-UDCA) is a non-volatile
derivative with high potentiality to be incorporated into nanocarriers. Therefore, in this work GER-UDCA-loaded
Solid Lipid Nanoparticles (SLNs) and PLGA nanoparticles (NPs) intended for nose-to-brain delivery were
developed and characterized. SLNs were prepared by emulsion/solvent evaporation method and NPs by
nanoprecipitation method. Briefly, formulations were optimized considering various processing variables and
nanoparticle characterization was performed in terms of morphology, size, surface charge, drug loading (DL%),
encapsulation efficiency (EE%) and in vitro drug release. Finally, the stability of free and encapsulated GERUDCA
was evaluated in enzymatic medium from rat liver homogenates. GER-UDCA-SLN and GER-UDCA-NPs
showed spherical shape, mean size of 120/180 nm with polydispersity index < 0.2, and zeta potential around
−22/-26 mV, respectively. After freeze-drying, the DL% was 6% for SLN and 12% for NPs with EE% values of
89.3% and 60.1%, respectively. Preliminary data regarding in vitro release of GER-UDCA from the nanoparticles
evidenced a higher dissolution rate than the free drug, probably due to the increase of surface contact. Results
in liver rat homogenate suggested a contribution of the nanoparticles in the stability of the prodrug in
physiologic environments. In conclusion, these GER-UDCA-loaded nanocarriers demonstrated a possible
application in further in vivo studies of nose-to-brain drug delivery.
2018
- Self-assembling organogelators for artificial stratum corneum models: key-role parameters in skin permeation prediction
[Poster]
Iannuccelli, V.; Maretti, E.; Rustichelli, C.; Miselli, P.; Truzzi, E.; Leo, E.
abstract
The development of in vitro methods to predict in vivo percutaneous absorption of bioactive molecules is a challenge to which the researchers are called in order to eliminate or reduce the pharmacological and toxicological tests on animal models. Artificial stratum corneum (SC) models obtained by self-assembled oganogelators were designed for skin permeation assessment of butyl methoxydibenzoylmethane (BMDBM, log Po/w = 4.68) and methylene blue (MB, log Po/w = 0.91). A multi-analytical approach was adopted to provide detailed understanding about the gelator organization within the models and find possible parameters playing a key-role in in vivo and ex vivo SC permeation prediction. The evaluation of in vitro skin permeation data compared with those obtained ex vivo and previously in vivo on humans for BMDBM showed good correlations vitro/ex vivo and vitro/vivo for both butyl BMDBM, as the lipophilic permeant, and MB, as the hydrophilic permeant, by using TS20 as well as both STS and ME models. With the aim of providing detailed understanding about the organogelator behaviour and organization within the models and find possible parameters playing a key-role in SC permeation prediction a multi-analytical approach was adopted. All the models did not flow upon tube tilting and could be described as gels, with the exception of STS10 model that appeared as thick liquid being gelator concentration lower than mgc value. Unlike SA and TS models that exhibited networks capable of immobilizing completely the solvent, STS and ME10 models revealed the syneresis phenomenon according to gelator concentration. The actual presence of water within STS aggregates (reverse micelles) of ME models was demonstrated by means of TG/DTA analysis showing two thermal events in the range of about 50-130°C related to removal of water molecules. Unlike the pure gelators, XRPD profiles from all the SC models exhibited a broad peak at about 20° 2θ indicating the presence of a networked structure of the gelators where the width of the peak at half maximum is dependent on the crystallinity of the sample, which in turn is dependent on non-covalent interactions amongst the gelator molecules responsible for the formation of an ordered structure. Intermolecular interactions also arisen from FT-IR spectra showing subsided ester group stretching in TS, STS, and ME models. Architectural arrangements of the organogelators within TS, STS, and ME models, as outlined by microscopy analyses, involved round or worm-like architectures of spherulitic clusters. Under polarized light, the occurrence of birefringence revealed the so-called “maltese crosses” in STS models that are characteristic of liquid crystals with lamellar structures. The results demonstrated the relevant role of both the arrangement of gelator packing and crystallinity extent in mimicking SC in vivo/ex vivo skin permeation of both lipophilic and hydrophilic compounds. These findings could account for the behaviour and development of other artificial skin models involving different materials for the skin permeation prediction.
2018
- Small-angle neutron scattering characterization of liposomes for anti-tuberculosis inhaled therapy.
[Poster]
Truzzi, Eleonora; Angela, Capocefalo; Fabio, Domenici; Carlo, Castellano; Fiorella, Meneghetti; Iannuccelli, Valentina; Maretti, Eleonora; Leo, Eliana Grazia; Costantino, Luca
abstract
The present investigation studied the effects of two first-line anti-tuberculosis (TB) drugs, rifampicin (RIF) and isoniazid (INH), on the structure of multilamellar liposomes. Liposomes have been shown to be a promising system for inhaled therapy. (1) The study of liposome-drug interactions is essential, and small-angle neutron scattering (SANS) technique provides valuable and unique data about steric bilayer thickness, particle dispersion, number of lamellae and drug localization under physiological conditions. (2) Unloaded, single drug-loaded and co-loaded liposomes were prepared using different amounts of drugs by reverse phase evaporation method. Liposomal suspensions were prepared using D2O, in order to emphasize the contrast between the aqueous and the lipid/drug phases. The samples were characterized by dynamic light scattering, atomic force microscopy and finally by SANS technique (Rutherford Appleton Laboratory, U.K.). Neutron scattering curves were analyzed using a multi-shell spherical model of the fitting routine SASView 2.2.0. Liposomes have been shown to be physico-chemically stable during the experiments, efficiently drug-loaded, and able to control drug release. Dimensional analysis demonstrated that particle sizes are in the range of SANS dimensional detection. SANS curves exhibited Bragg peaks for all samples, confirming the multilamellar liposome structure. By fitting the data, significant differences among the samples have been highlighted. RIF-liposomes were less ordered than unloaded liposomes: a reduction of the lamellae number was observed and the periodicity of the lipid bilayers slightly increased with the increment of the drug loading, may be due to RIF interaction with phospholipid tails, which can destabilize liposome lamellarity, since RIF is a hydrophobic drug. In INH-liposomes, the drug payloads did not change vesicle structure, because INH is a hydrophilic drug. However, INH induced a change in the inter-bilayer periodical spacing, which could be compatible with the formation of drug-liposome complexes at the water-lipid interface. Finally, the RIF-INH co-loaded liposomes exhibited the same characteristics of unloaded liposomes, suggesting that INH and RIF together have a stabilizing effect on the structure. In fact, no destabilization and no changing in inter-bilayer periodical spacing were observed. In conclusion, SANS analysis provides fundamental information about drug-liposome interactions to comprehend the relation between system structure behaviour and its biological activity. Moreover, data suggest that the co-encapsulation of the two anti-TB drugs may have a synergic effect on liposome stability.
1. A. Elhissi, Curr. Pharm. Des., 2017, 23, 362-372.
2. PC. Lin, S. Lin, PC. Wang, R. Sridhar, Biotechnol. Adv., 2014, 32, 711– 726.
2017
- DEVELOPMENT AND COMPARISON OF TWO LIPID-BASED NANOSYSTEMS FOR THE CO-DELIVERY OF RIFAMPICIN AND ISONIAZID: SMALL-ANGLE NEUTRON SCATTERING CHARACTERISATION
[Abstract in Atti di Convegno]
Truzzi, Eleonora; Maretti, Eleonora; Meneghetti, Fiorella; Castellano, Carlo; Iannuccelli, Valentina; Leo, Eliana
abstract
Recently lipid-based drug delivery system (DDS), such as Solid Lipid Nanoparticles (SLNs) and liposomes, have been proposed in the inhaled therapy to improve the drug targeting and delivery.
In this work SLNs and liposomes were studied for the co-delivery of two first-line anti-tuberculosis drugs, Rifampicin (RIF) and Isoniazid (INH). RIF and INH were chosen as model drugs, since they display different physicochemical features: RIF is a poor water soluble compound, while INH is extremely hydrophilic.
Both lipid-based DDS (SLN and liposomes) have been characterized regarding size and zeta potential by DSL (dynamic light scattering) and regarding shape and external morphology by AFM (atomic force microscopy) and by SEM (scanning electron microscopy).
Moreover, drug content, in vitro drug release and stability in suspension, were evaluated in order to highlight the effects of the co-encapsulation. In fact, drugs can interact with the delivery systems modifying its properties and in vitro behaviour. In order to comprehend and evaluate the possible interactions between drugs and DDS, loaded SLNs and liposomes has been further studied by small-angle neutron scattering (SANS).
Small-Angle Neutron Scattering (SANS) is a useful tool in gaining a detailed understanding of the structure of macromolecular systems that make up SLN and liposomes. SANS is widely applicable, with the inherent advantage over visual techniques, of probing accurately structural features on nanometer length scales as well as yielding ensemble-averaged information on freely water-suspended membranes. Of note, the SANS D2O/H2O contrast variation methodology allows a well distinguished scattering from the shell with respect to the corresponding inner content. Besides, SANS do not entail the addition or inclusion of bulky fluorescent dyes or labels needed by other techniques (i.e. fluorescence and electron microscopy), which can perturb or affect the phase behavior observed.
2017
- Nuove molecole funzionalizzanti per il targeting ai macrofagi di vettori lipidici
[Brevetto]
Iannuccelli, Valentina; Maretti, Eleonora; Costantino, Luca; Leo, Eliana; Rustichelli, Cecilia; Truzzi, Eleonora
abstract
La presente invenzione ha per oggetto molecole derivate dal mannosio ed
il loro uso per direzionare carrier farmacologici in maniera specifica verso i
macrofagi. L’invenzione consiste nella sintesi di nuove molecole derivate
dal mannosio utilizzate come funzionalizzanti di superficie per vettori
micro- e nano- particellari progettati per attività intra-5 macrofagica di
farmaci somministrabili per via inalatoria.
2017
- Self-Assembled Lipid Nanoparticles for Oral Delivery of Heparin-Coated Iron Oxide Nanoparticles for Theranostic Purposes
[Articolo su rivista]
Truzzi, Eleonora; Bongio, Chiara; Sacchetti, Francesca; Maretti, Eleonora; Montanari, Monica; Iannuccelli, Valentina; Vismara, Elena; Leo, Eliana Grazia
abstract
Recently, solid lipid nanoparticles (SLNs) have attracted increasing attention owing to their potential as an oral delivery system, promoting intestinal absorption in the lymphatic circulation which plays a role in disseminating metastatic cancer cells and infectious agents throughout the body. SLN features can be exploited for the oral delivery of theranostics. Therefore, the aim of this work was to design and characterise self-assembled lipid nanoparticles (SALNs) to encapsulate and stabilise iron oxide nanoparticles non-covalently coated with heparin (Fe@hepa) as a model of a theranostic tool. SALNs were characterised for physico-chemical properties (particle size, surface charge, encapsulation efficiency, in vitro stability, and heparin leakage), as well as in vitro cytotoxicity by methyl thiazole tetrazolium (MTT) assay and cell internalisation in CaCo-2, a cell line model used as an indirect indication of intestinal lymphatic absorption. SALNs of about 180 nm, which are stable in suspension and have a high encapsulation efficiency (>90%) were obtained. SALNs were able to stabilise the heparin coating of Fe@hepa, which are typically unstable in physiological environments. Moreover, SALNs-Fe@hepa showed no cytotoxicity, although their ability to be internalised into CaCo-2 cells was highlighted by confocal microscopy analysis. Therefore, the results indicated that SALNs can be considered as a promising tool to orally deliver theranostic Fe@hepa into the lymphatic circulation, although further in vivo studies are needed to comprehend further potential applications.
2017
- Surface engineering of Solid Lipid Nanoparticle assemblies by methyl α-d-mannopyranoside for the active targeting to macrophages in anti-tuberculosis inhalation therapy
[Articolo su rivista]
Maretti, Eleonora; Costantino, Luca; Rustichelli, Cecilia; Leo, Eliana Grazia; Croce, Maria Antonietta; Francesca, Buttini; Truzzi, Eleonora; Iannuccelli, Valentina
abstract
This study describes the development of new mannosylated Solid Lipid Nanoparticle assemblies (SLNas) delivering rifampicin for an inhaled treatment of tuberculosis. SLNas were surface engineered with mannose residues to recognize mannose receptors located on infected alveolar macrophages and facilitate cell internalization. Two sets of SLNas were produced by the melt emulsifying technique using biocompatible lipid components, i.e. cholesteryl myristate combined with palmitic acid (PA set) or tripalmitin (TP set), in the presence of the targeting moiety, methyl α-d-mannopyranoside. Mannosylated SLNas were examined for their physical properties, drug payloads and release, as well as respirability in terms of emitted dose and respirable fraction determined by Next Generation Impactor. The most appropriate formulations were assessed for mannosylation using FTIR, XPS, SEM coupled with EDX analysis, and wettability assay, in comparison with the respective non-functionalized SLNas. Besides, cytotoxicity and cell internalization ability were established on J774 murine macrophage cell line. Mannosylated SLNas exhibited physical properties suitable for alveolar macrophage passive targeting, adequate rifampicin payloads (10-15%), and feasible drug maintenance within SLNas along the respiratory tract before macrophage internalization. Despite respirability impaired by powder cohesiveness, surface mannosylation provided quicker macrophage phagocytosis, giving evidence of an active targeting promotion.
2016
- Anti-TB inhalation therapy: Design of mannose-based functionalised Solid Lipid Microparticles for an active targeting to alveolar macrophages
[Abstract in Atti di Convegno]
Maretti, Eleonora; Rustichelli, Cecilia; Costantino, Luca; Truzzi, Eleonora; Sacchetti, Francesca; Leo, Eliana Grazia; Iannuccelli, Valentina
abstract
Human tuberculosis (TB) is mainly a disease of the lung characterised by a long chronic stage of infection and progressive pathology that compromise the respiratory system. This is a curable infectious bacterial disease caused by the Mycobacterium tuberculosis (Mtb). TB therapies have exploited conventional routes of administration, such as oral and intramuscular1. The pulmonary route appears the most reasonable and effective way to target the alveolar macrophages (AM) and eradicate surviving Mtb at the primary infected site of TB, especially considering that 75-80% of cases remain localised in the lungs. The anti-TB therapy by inhalation offers benefits compared with the current treatment in terms of patient’s compliance improvement, reduction in dose amount and frequency, treatment duration and TB diffusion in other organs, thus minimising the risk of drug-resistant mutants, toxicity and side effects.
For a direct intramacrophagic antitubercular therapy using Dry Powder Inhaler (DPI) devices, Solid Lipid Microparticles (SLM), produced using the melt emulsifying technique followed by freeze-drying, were developed to load rifampicin, a first-line antitubercular drug.
In the present project, SLM were modified to improve drug loading level and release as well as AM targeting. Several biocompatible lipid components such as fatty acids and their derivatives, diglycerides and triglycerides, were processed using mixtures of biocompatible stabilisers (sodium taurocholate and methyl mannopyranoside) in order to obtain SLM with maximum efficiency in terms of drug loading and release in simulated lung fluid. Lipids in the liquid physical state embedded into SLM provided Microstructured Lipid Carriers (MLC) that are known to exhibit superior advantages over SLM such as enhanced drug loading capacity and prevention of drug expulsion intended to maximise the drug concentration at the primary site of TB infection. The obtained microcarriers were examined for their intrinsic properties such as size and size distribution, morphology and shape, surface charge, bulk and tap density, aerodynamic diameter, physical state of the components, wettability, drug loading and release.
Macrophages, as is common knowledge, possess mannose-specific membrane receptors (MR) that can be recognised by carriers bearing mannose residues, facilitating their internalisation 2, 3.
Therefore, the functionalisation of SLM surface by mannose derivatives used as the co- stabiliser in the SLM formulation was used to achieve an active targeting. The actual presence of mannose on SLM surface was investigated by means of X-ray Photoelectron Spectroscopy for Chemical Analysis (XPS) and Energy Dispersive X-ray Analysis (EDX).
2016
- Characterization of natural clays from italian deposits with focus on elemental composition and exchange estimated by edx analysis: potential pharmaceutical and cosmetic uses
[Articolo su rivista]
Iannuccelli, Valentina; Maretti, Eleonora; Sacchetti, Francesca; Romagnoli, Marcello; Bellini, Alessia; Truzzi, Eleonora; Miselli, Paola; Leo, Eliana Grazia
abstract
Purification processes performed on natural clays to select specific clay minerals are complex
and expensive and can lead to over-exploitation of some deposits. The present study aimed to examine
physicochemical (mineralogy, morphology, size, surface charge, chemical composition, cation exchange
capacity [CEC], and pH) and hydration (swelling, wettability, water sorption, and rheological behavior)
properties of three native clays from Italian deposits for potential pharmaceutical and cosmetic uses due to
the presence of phyllosilicate minerals. Particular emphasis was placed on energy dispersive X-ray (EDX)
microanalysis coupled with the ‘cesium method’ to assay clay elemental composition and CEC. One
bentonite of volcanic origin (BNT) and two kaolins, one of hydrothermal origin (K-H) and another of
lacustrine-fluvial origin (K-L), were evaluated in comparison with a commercial, purified bentonite. The
CEC assay revealed the complete substitution of exchangeable cations (Na+ and Ca2+) by Cs+ in BNT
samples and CEC values consistent with those of typical smectites (100.64 7.33 meq/100 g). For kaolins,
partial substitution of Na+ cations occurred only in the K-L samples because of the interstratified mineral
component which has small CEC values (11.13 5.46 meq/100 g for the K-H sample and 14.75
6.58 meq/100 g for the K-L sample). The degree of isomorphous substitution of Al3+ by Mg2+ affected the
hydration properties of BNT in terms of swelling, water sorption, and rheology, whereas both of the poorly
expandable kaolins exhibited significant water-adsorption properties. The EDX microanalysis has proved
to be of considerable interest in terms of providing more information about clay properties in comparison
with other commonly used methods and to identify the role played by both chemical and mineralogical
composition of natural clays for their appropriate use in pharmaceutical and cosmetic fields.
2016
- Development and in vitro characterization of SLN encapsulating magnetic Heparin coated Iron Oxide for theranostic application
[Abstract in Atti di Convegno]
Truzzi, Eleonora; Mazza, Federica; Sacchetti, Francesca; Maretti, Eleonora; Iannuccelli, Valentina; Vismara, Elena; Leo, Eliana Grazia
abstract
Solid Lipid Nanoparticles (SLN) have been proposed for the oral delivery of drugs with poor oral bioavailability for their ability to be internalized directly by the lymphatic circulation, like chylomicrons, through intestinal absorption. Lymphatic system is considered an interesting target for anti-cancer drugs and contrast agents because exerts an active role in the cancer metastasis being the major route of the solid tumor spread. Recently, according to the potentiality of the iron oxide in diagnostic and of heparin in the cancer therapy, iron oxide nanoparticles non-covalently coated with heparin (Fe@hepa) have been proposed as delivery systems for theranostic application (1). The aim of this work was to encapsulateFe@hepa in a biocompatible solid lipid shell in order to obtain a nanotheranostic tool for promoting the oral absorption through the lymphatic route. SLN have been formulated by a modified self nanoemulsifying technique by using Gelucire50/13 and Geleol™. The resulted Fe@hepa-SLN were characterized regarding size, morphology, storage stability as well in vitro release of heparin and iron oxide. In addition, preliminary studies on Caco-2 cell line were carried out evaluating cytotoxicity by MTT tests and internalization by the direct quantification of Fe@hepa inside the cells. Fe@hepa-SLN displayed a mean diameter below 300 nm, suitable for the oral administration, and an incorporation efficiency of 75% ± 3.9. Morphology analyisis showed the lipid shell surrounding the Fe@hepa nanoparticles and the release studies demonstrated that this lipid envelop stabilized the heparin coating in physiological conditions. Finally, studies on Caco2 cells showed the low cytotoxicity of the Fe@hepa-SLN and their ability to be internalized in the cells used as intestinal permeability model. These results indicate that this novel nanotechnology strategy could be a promising tool for oral nanotheranostic approaches.
2016
- Medicinal Chemistry drives drug targeted delivery: a successful interplay
[Abstract in Atti di Convegno]
Costantino, Luca; Maretti, Eleonora; Rustichelli, Cecilia; Truzzi, Eleonora; Sacchetti, Francesca; Leo, Eliana Grazia; Iannuccelli, Valentina
abstract
Targeted drug delivery is object of an intense research. A medicinal chemistry approach allowed us to modify an opioid peptide in order to remove the opioid activity and retain the ability to cross the blood-brain barrier. Polyester-based nanoparticles (Np) surface-decorated with this peptide were shown to be able to deliver loperamide (a model drug) into CNS, as well as cholesterol (for the treatment of Huntington’s disease) and albumin (a model of a cargo protein). With a view of clinical translation, however, polyester Np seems to be not well suited for CNS diseases. Thus, we decided to use our peptide in alternative ways other than as a conjugate with the carboxyl group of the polyester PLGA, starting material for the production of the Np. We develop a new synthetic procedure that allow the conjugation of the peptide with substrates containing hydroxyl groups, less reactive than the carboxy group of the polyester (we considered as a substrate the poly(vinyl alcohol)), as well as a linker that could be inserted between a cargo and the peptide targeting moiety. At the same time, we moved towards lipidic carriers, a kind of delivery agents already clinically available. To gain experience on these carriers, we designed rifampicin-loaded Solid Lipid Nanoparticle assemblies (SLNas), for a direct intramacrophagic antitubercular therapy using Dry Powder Inhaler (DPI) devices and, in the first instance, methyl mannopyranoside as targeting ligand, able to interact with mannose receptors present on macrophages. Results obtained will be presented and discussed.
2016
- SURFACE ENGINEERING OF SOLID LIPID NANOASSEMBLIES FOR INHALED INTRAMACROPHAGIC ANTI-TB THERAPY
[Abstract in Atti di Convegno]
Costantino, Luca; Maretti, Eleonora; Truzzi, Eleonora; Rustichelli, Cecilia; Leo, Eliana Grazia; Zapparoli, Mauro; Iannuccelli, Valentina
abstract
For an inhaled tuberculosis (TB) treatment, antibiotic aerosolization has to be produced by using drugs in their solid state administered by means of Dry Powder Inhaler (DPI) devices. In this regard, untreated drugs generally fail to reach alveolar epithelium and penetrate alveolar macrophages (AM) as the primary site of the infection.1 Therefore, the urgency to treat TB disease effectively may be addressed with approaches consisting of micro- or nanoparticulate carriers redeveloping existing drugs to reach the intended goal.2, 3 Specific modifications of the particulate carrier surface by conjugation with molecules that can specifically bind the receptors (active targeting) are expected to boost the particle avidity to cells increasing accumulation and intracellular uptake. Macrophages possess mannose-specific membrane receptors (MR) that can recognize and facilitate the internalization of carriers bearing mannose residues. In particular, the infected AM have an overexpression of MR.4 In the present study, surface engineered Solid Lipid Nanoparticle assemblies (SLNas) were developed as potential carriers of rifampicin, a first choice antitubercular drug, intended to maximize drug concentration at the primary site of TB infection. To increase specificity for macrophages and internalization potential, SLNas surface was functionalized by a mannosylated derivative to induce AM active targeting. Biocompatible lipid components such as fatty acids and their derivatives, diglycerides and triglycerides were processed by means of the melt emulsifying technique using biocompatible surfactants (sodium taurocholate and methyl mannopyranoside). Mannosylated SLNas were examined for their intrinsic properties (size and size distribution, shape, surface charge, bulk and tap density, aerodynamic diameter, porosity, flowability, physical state of the components). Powder breathability in terms of Emitted Dose and Fine Particle Fraction was assayed by Next Generation Impactor (NGI). This information on powder interparticle adhesion and deaggregation ability influencing powder dispersion and deposition onto alveolar epithelia. SLNas mannosylation was investigated by means of X-ray Photoelectron Spectroscopy for Chemical Analysis and Energy Dispersive X-ray Analysis. Prototypes of SLNas in terms of successful functionalization, optimal breathability and chemico-physical stability, were examined for cytotoxicity by MTT test on murine macrophages J774 cell lines.